Real-Time Detection of Polymerase Activity Using Supercritical Angle Fluorescence |
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| Authors: | Alexander Krieg Thomas Ruckstuhl Stephan Laib Stefan Seeger |
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| Affiliation: | 1. Physikalisch-Chemisches Institut, Universit?t Zürich, Winterthurerstr. 190, 8057, Zürich, Switzerland
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| Abstract: | We investigated the incorporation efficiencies of different fluorescently labelled dNTPs with polymerases by complementary strand synthesis. For this reason single stranded DNA was immobilized on a coverslip and the increase of fluorescence due to the synthesis of the corresponding strand with tagged dNTPs was detected with a supercritical angle fluorescence biosensor in real-time. By comparison of the observed signal intensities it was possible to conclude that the system Cy5-dCTP-Klenow (exonuclease free) fragment gives the best incorporation yield of the investigated enzymes and dNTPs. |
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| Keywords: | Biosensors fluorescence dyes polymerase primer elongation |
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